bims-glecem 2022-02-20 papers

bims-glecem

Biomed News

on Glycogen metabolism in exercise, cancer and energy metabolism

Issue of 2022–02–20
24 papers selected by
Dipsikha Biswas, Københavns Universitet

GSK3β and the aging kidney.
Regulation of Autophagy by the Glycogen Synthase Kinase-3 (GSK-3) Signaling Pathway.
An optimized method for tissue glycogen quantification.
Effect of 12-wk Training in Ovariectomised Rats on PGC-1α, NRF-1 and Energy Substrates.
Isoform-Specific Role of GSK-3 in High Fat Diet Induced Obesity and Glucose Intolerance.
Glycogen Synthase Kinase 3β Involvement in Neuroinflammation and Neurodegenerative Diseases.
Propolin G-Suppressed Epithelial-to-Mesenchymal Transition in Triple-Negative Breast Cancer Cells via Glycogen Synthase Kinase 3β-Mediated Snail and HDAC6-Regulated Vimentin Degradation.
TDAG51-Deficiency Podocytes are Protected from High-Glucose-Induced Damage Through Nrf2 Activation via the AKT-GSK-3β Pathway.
Indirubin-3'-alkoxime derivatives for upregulation of Wnt signaling through dual inhibition of GSK-3β and the CXXC5-Dvl interaction.
Age-related GSK3β overexpression drives podocyte senescence and glomerular aging.
A non-catalytic scaffolding activity of hexokinase 2 contributes to EMT and metastasis.
Characterization of a cold-adapted debranching enzyme and its role in glycogen metabolism and virulence of Vibrio vulnificus MO6-24/O.
1-(7-Chloroquinolin-4-yl)-N-(4-Methoxybenzyl)-5-Methyl-1H-1,2, 3-Triazole-4- carboxamide Reduces Aβ Formation and Tau Phosphorylation in Cellular Models of Alzheimer's Disease.
Multi-target neuroprotective effects of herbal medicines for Alzheimer's disease.
Magnesium-Assisted Cisplatin Inhibits Bladder Cancer Cell Survival by Modulating Wnt/β-Catenin Signaling Pathway.
The GSK-3 Inhibitor CT99021 Enhances the Acquisition of Spatial Learning and the Accuracy of Spatial Memory.
Energy metabolism response of Litopenaeus vannamei to combined stress of acute cold exposure and waterless duration: Implications for physiological regulation and waterless live transport.
Effects of Perch Essence Supplementation on Improving Exercise Performance and Anti-Fatigue in Mice.
GSK-3β Localizes to the Cardiac Z-Disc to Maintain Length Dependent Activation.
Tideglusib Ameliorates Ischemia/Reperfusion Damage by Inhibiting GSK-3β and Apoptosis in Rat Model of Ischemic Stroke.
Physiological Function during Exercise and Environmental Stress in Humans-An Integrative View of Body Systems and Homeostasis.
GBE1 Is an Independent Prognostic Marker and Associated With CD163+ Tumor-Associated Macrophage Infiltration in Lung Adenocarcinoma.
HIGH PREVALENCE OF PARASPINAL MUSCLE INVOLVEMENT IN ADULTS WITH McARDLE DISEASE.
Triggering the nanophase separation of albumin through multivalent binding to glycogen for drug delivery in 2D and 3D multicellular constructs.

J Clin Invest. 2022 02 15. pii: e155885. [Epub ahead of print]132(4):

GSK3β and the aging kidney.

Jordan A Kreidberg, Valerie A Schumacher.

Kidney function decreases with age and may soon limit millions of lives as the proportion of the population over 70 years of age increases. Glycogen synthase kinase 3β (GSK3β) is involved with metabolism and may have a role in kidney senescence, positioning it as a target for complications from chronic kidney disease. However, different studies suggest GSK3 has contrasting effects. In this issue of the JCI, Fang et al. explored the function of GSK3β and the interplay with lithium using human tissue and mouse models. Notably, GSK3β was overexpressed and activated in aging mice, and depleting GSK3β reduced senescence and glomerular aging. In this Commentary, we explore the similarities and differences between Fang et al. and previous findings by Hurcombe et al. These findings should prompt further study of lithium and other GSK3β inhibitors as a means of extending glomerular function in individuals with chronic kidney disease.

DOI: https://doi.org/10.1172/JCI155885
Int J Mol Sci. 2022 Feb 01. pii: 1709. [Epub ahead of print]23(3):

Regulation of Autophagy by the Glycogen Synthase Kinase-3 (GSK-3) Signaling Pathway.

Hsuan-Yeh Pan, Mallika Valapala.

  Autophagy is a vital cellular mechanism that benefits cellular maintenance and survival during cell stress. It can eliminate damaged or long-lived organelles and improperly folded proteins to maintain cellular homeostasis, development, and differentiation. Impaired autophagy is associated with several diseases such as cancer, neurodegenerative diseases, and age-related macular degeneration (AMD). Several signaling pathways are associated with the regulation of the autophagy pathway. The glycogen synthase kinase-3 signaling pathway was reported to regulate the autophagy pathway. In this review, we will discuss the mechanisms by which the GSK-3 signaling pathway regulates autophagy. Autophagy and lysosomal function are regulated by transcription factor EB (TFEB). GSK-3 was shown to be involved in the regulation of TFEB nuclear expression in an mTORC1-dependent manner. In addition to mTORC1, GSK-3β also regulates TFEB via the protein kinase C (PKC) and the eukaryotic translation initiation factor 4A-3 (eIF4A3) signaling pathways. In addition to TFEB, we will also discuss the mechanisms by which the GSK-3 signaling pathway regulates autophagy by modulating other signaling molecules and autophagy inducers including, mTORC1, AKT and ULK1. In summary, this review provides a comprehensive understanding of the role of the GSK-3 signaling pathway in the regulation of autophagy.

Keywords:  AKT; GSK-3; GSK-3β; PKC; TFEB; ULK1; autophagy; lysosome; mTORC1

DOI:  https://doi.org/10.3390/ijms23031709
Physiol Rep. 2022 Feb;10(4): e15195

An optimized method for tissue glycogen quantification.

Kyle J Schaubroeck, Brooks P Leitner, Rachel J Perry.

Mobilization of glycogen, the short-term storage form of glucose, is the body's first defense against hypoglycemia and is critical for energy provision during high intensity exercise. Therefore, to advance metabolic research, it is critical to be able to accurately measure glycogen concentrations, including during a prolonged fast and other glycogen-modulating interventions. Unfortunately, prior enzymatic methods of glycogen detection have been plagued by poor detection in the lower range, high sample mass requirements, and complicated and/or expensive protocols which introduce substantial technical variability into the measured glycogen concentrations. To address these limitations, here we report a streamlined and versatile glycogen extraction protocol coupled with an optimized phenol-sulfuric acid quantification protocol. With this method, we demonstrate how glycogen can be extracted from only 20 mg of tissue with one centrifugation step and quantified with a highly precise (Intra-assay %CV ranges from 5-10%) and sensitive (proportionality constant for glycogen = 0.07279 A.U./µg) assay. The cost of all materials equates to ~10 cents per sample. Therefore, this method represents an attractive means of assessing ex vivo tissue glycogen content including at the extremes of glycogen concentrations.

DOI: https://doi.org/10.14814/phy2.15195
Int J Sports Med. 2022 Feb 18.

Effect of 12-wk Training in Ovariectomised Rats on PGC-1α, NRF-1 and Energy Substrates.

Taciane Maria Melges Pejon, Vinicius Silva Faria, Claudio Alexandre Gobatto, Fúlvia Barros Manchado-Gobatto, PedroPauloMenezes Scariot, AnabelleSilva Cornachione, Wladimir Rafael Beck.

Metabolic diseases are associated with hypoestrogenism owing to their lower energy expenditure and consequent imbalance. Physical training promotes energy expenditure through PGC-1α and NRF-1, which are muscle proteins of the oxidative metabolism. However, the influence of physical training on protein expression in individuals with hypoestrogenism remains uncertain. Thus, the aim of this study is to determine the effect of 12 weeks of moderate-intensity swimming training on the muscle expression of PGC-1α, NRF-1, glycogen and triglyceride in ovariectomised rats. OVX and OVX+TR rats were subjected to ovariectomy. The trained animals swam for 30 minutes, 5 days/week, at 80% of the critical load intensity. Soleus was collected to quantify PGC-1α and NRF-1 expressions, while gastrocnemius and gluteus maximus were collected to measure glycogen and triglyceride. Blood glucose was also evaluated. Whereas ovariectomy decreased PGC-1α expression (p<0.05) without altering NRF-1 (p=0.48), physical training increased PGC-1α (p<0.01) and NRF-1 (p<0.05). Ovariectomy reduced glycogen (p<0.05) and triglyceride (p<0.05), whereas physical training increased glycogen (p<0.05) but did not change triglyceride (p=0.06). Ovariectomy increased blood glucose (p<0.01), while physical training reduced it (p<0.01). In summary, 12 weeks of individualized and moderate-intensity training were capable of preventing muscle metabolic consequences caused by ovariectomy.

DOI: https://doi.org/10.1055/a-1717-1693
Cells. 2022 Feb 05. pii: 559. [Epub ahead of print]11(3):

Isoform-Specific Role of GSK-3 in High Fat Diet Induced Obesity and Glucose Intolerance.

Manisha Gupte, Sultan Tousif, Jacob J Lemon, Angelica Toro Cora, Prachi Umbarkar, Hind Lal.

  Obesity-associated metabolic disorders are rising to pandemic proportions; hence, there is an urgent need to identify underlying molecular mechanisms. Glycogen synthase kinase-3 (GSK-3) signaling is highly implicated in metabolic diseases. Furthermore, GSK-3 expression and activity are increased in Type 2 diabetes patients. However, the isoform-specific role of GSK-3 in obesity and glucose intolerance is unclear. Pharmacological GSK-3 inhibitors are not isoform-specific, and tissue-specific genetic models are of limited value to predict the clinical outcome of systemic inhibiion. To overcome these limitations, we created novel mouse models of ROSA26CreERT2-driven, tamoxifen-inducible conditional deletion of GSK-3 that allowed us to delete the gene globally in an isoform-specific and temporal manner. Isoform-specific GSK-3 KOs and littermate controls were subjected to a 16-week high-fat diet (HFD) protocol. On an HFD, GSK-3α KO mice had a significantly lower body weight and modest improvement in glucose tolerance compared to their littermate controls. In contrast, GSK-3β-deletion-mediated improved glucose tolerance was evident much earlier in the timeline and extended up to 12 weeks post-HFD. However, this protective effect weakened after chronic HFD (16 weeks) when GSK-3β KO mice had a significantly higher body weight compared to controls. Importantly, GSK-3β KO mice on a control diet maintained significant improvement in glucose tolerance even after 16 weeks. In summary, our novel mouse models allowed us to delineate the isoform-specific role of GSK-3 in obesity and glucose tolerance. From a translational perspective, our findings underscore the importance of maintaining a healthy weight in patients receiving lithium therapy, which is thought to work by GSK-3 inhibition mechanisms.

Keywords:  GSK-3; glucose tolerance; high-fat diet; obesity

DOI:  https://doi.org/10.3390/cells11030559
Curr Med Chem. 2022 Feb 16.

Glycogen Synthase Kinase 3β Involvement in Neuroinflammation and Neurodegenerative Diseases.

Teresa Gianferrara, Eleonora Cescon, Ilenia Grieco, Giampiero Spalluto, Stephanie Federico.

   BACKGROUND: GSK-3β activity has been strictly related to neuroinflammation and neurodegeneration. Alzheimer's disease is the most studied neurodegenerative disease, but GSK-3β seems to be involved in almost all neurodegenerative diseases including Parkinson's disease, amyotrophic lateral sclerosis, frontotemporal dementia, Huntington's disease and the autoimmune disease multiple sclerosis.
OBJECTIVE: The aim of this review is to help researchers both working on this research topic or not to have a comprehensive overview on GSK-3β in the context of neuroinflammation and neurodegeneration.
METHOD: Literature has been searched using PubMed and SciFinder databases by inserting specific keywords. A total of more than 500 articles have been discussed.
RESULTS: First of all, the structure and regulation of the kinase were briefly discussed and then, specific GSK-3β implications in neuroinflammation and neurodegenerative diseases were illustrated also with the help of figures, to conclude with a comprehensive overview on the most important GSK-3β and multitarget inhibitors. For all discussed compounds, the structure and IC50 values at the target kinase have been reported.
CONCLUSION: GSK-3β is involved in several signaling pathways both in neurons as well as in glial cells and immune cells. The fine regulation and interconnection of all these pathways are at the base of the rationale use of GSK-3β inhibitors in neuroinflammation and neurodegeneration. In fact, some compounds are now under clinical trials. Despite this, pharmacodynamic and ADME/Tox profiles of the compounds were often not fully characterized and this is deleterious in such a complex system.

Keywords:  Alzheimer's disease; GSK-3β inhibitors; Glycogen Synthase Kinase 3β; Multiple Sclerosis; Parkinson's disease; multitarget ligands; neurodegeneration; neuroinflammation

DOI:  https://doi.org/10.2174/0929867329666220216113517
Int J Mol Sci. 2022 Jan 31. pii: 1672. [Epub ahead of print]23(3):

Propolin G-Suppressed Epithelial-to-Mesenchymal Transition in Triple-Negative Breast Cancer Cells via Glycogen Synthase Kinase 3β-Mediated Snail and HDAC6-Regulated Vimentin Degradation.

Jih-Tung Pai, Xing-Han Chen, Yann-Lii Leu, Meng-Shih Weng.

  Triple-negative breast cancer (TNBC) is a highly aggressive breast cancer with a poor prognosis. The incidence and mortality rate of TNBC are frequently found in younger women. Due to the absence of a good therapeutic strategy, effective remedies for inhibiting TNBC have been developed for improving the cure rate. Epithelial-to-mesenchymal transition (EMT) is a critical mechanism to regulate cancer cell motility and invasion. Furthermore, ectopic expression of EMT molecules correlates with the metastasis and poor prognosis of TNBC. Targeting EMT might be a strategy for the therapy and prevention of TNBC. Propolin G, an active c-prenylflavanone in Taiwanese propolis, has been shown to possess anti-cancer activity in many cancers. However, the anti-metastasis activity of propolin G on TNBC is still unclear. The present study showed that the migration and invasion activities of TNBC cells was suppressed by propolin G. Down-regulated expression of Snail and vimentin and up-regulated expression of E-cadherin were dose- and time-dependently observed in propolin G-treated MDA-MB-231 cells. Propolin G inhibited Snail and vimentin expressions via the signaling pathways associated with post-translational modification. The activation of glycogen synthase kinase 3β (GSK-3β) by propolin G resulted in increasing GSK-3β interaction with Snail. Consequently, the nuclear localization and stability of Snail was disrupted resulting in promoting the degradation. Propolin G-inhibited Snail expression and the activities of migration and invasion were reversed by GSK-3β inhibitor pretreatment. Meanwhile, the outcomes also revealed that histone deacetylase 6 (HDAC6) activity was dose-dependently suppressed by propolin G. Correspondently, the amounts of acetyl-α-tubulin, a down-stream substrate of HDAC6, were increased. Dissociation of HDAC6/Hsp90 with vimentin leading to increased vimentin acetylation and degradation was perceived in the cells with the addition of propolin G. Moreover, up-regulated expression of acetyl-α-tubulin by propolin G was attenuated by HDAC6 overexpression. On the contrary, down-regulated expression of vimentin, cell migration and invasion by propolin G were overturned by HDAC6 overexpression. Conclusively, restraint cell migration and invasion of TNBC by propolin G were activated by the expression of GSK-3β-suppressed Snail and the interruption of HDAC6-mediated vimentin protein stability. Aiming at EMT, propolin G might be a potential candidate for TNBC therapy.

Keywords:  (GSK-3β); epithelial-to-mesenchymal transition (EMT); glycogen synthase kinase 3β; histone deacetylase 6 (HDAC6); propolin G; triple-negative breast cancer (TNBC)

DOI:  https://doi.org/10.3390/ijms23031672
Inflammation. 2022 Feb 17.

TDAG51-Deficiency Podocytes are Protected from High-Glucose-Induced Damage Through Nrf2 Activation via the AKT-GSK-3β Pathway.

Chuntian Liu, Yanling Li, Xiaojuan Wang.

  T cell death-associated gene 51 (TDAG51) has been implicated in the development of various pathological conditions. However, whether TDAG51 plays a role in diabetic renal disease remains unknown. The current work investigated the possible function of TDAG51 in diabetic renal disease using high-glucose (HG)-stimulated podocytes in vitro. The elevation of TDAG51 was observed in podocytes in response to HG exposure and the glomeruli of diabetic mice. The siRNAs targeting TDAG51 were applied to deplete TDAG51 in HG-stimulated podocytes. Crucially, TDAG51 deficiency was sufficient to decrease the apoptosis, oxidative stress, and inflammation caused by HG. Mechanically, the inhibition of TDAG51 was capable of enhancing the activation of nuclear factor E2-related factor 2 (Nrf2) associated with the upregulation of AKT-glycogen synthase kinase-3β (GSK-3β) pathway. The reduction of AKT abolished the activation of Nrf2 elicited by TDAG51 deficiency. Additionally, the reduction of Nrf2 diminished the anti-HG injury effect elicited by TDAG51 deficiency. Overall, these data demonstrate that TDAG51 deficiency defends against HG-induced podocyte damage through Nrf2 activation by regulating AKT-GSK-3β pathway. This study suggests that TDAG1 may have a potential role in diabetic renal disease by affecting HG-induced podocyte damage.

Keywords:  Nrf2; TDAG51; diabetic renal disease; high glucose

DOI:  https://doi.org/10.1007/s10753-022-01638-9
Bioorg Chem. 2022 Feb 08. pii: S0045-2068(22)00069-4. [Epub ahead of print]121 105664

Indirubin-3'-alkoxime derivatives for upregulation of Wnt signaling through dual inhibition of GSK-3β and the CXXC5-Dvl interaction.

Doona Song, Yunja Lee, Min-Jeong Kang, Jae Won Kim, Soung-Hoon Lee, Kang-Yell Choi, Eun-Yeong Kim, Kiho Lee, Gyoonhee Han.

  Glycogen synthase kinase-3β (GSK-3β) appears to be ordinarily expressed, and functionally redundant in Wnt/β-catenin signaling. The Wnt proteins induce transduction of a cytoplasmic protein, Dishevelled (Dvl) which negatively modulates GSK-3β activity. CXXC5 is a negative modulator of the Wnt/β-catenin signaling through the interaction with Dvl in the cytosol. This indicates that Wnt/β-catenin signaling could be efficiently modulated by controlling GSK-3β and the CXXC5-Dvl interaction. In this study, we designed a series of indirubin-3'-oxime and indirubin-3'-alkoxime derivatives containing various functional groups at the 5- or 6-position (R1) alongside alkyl or benzylic moieties at the 3'-oxime position (R2). These activate Wnt signaling through inhibitions of both GSK-3β and the CXXC5-Dvl protein-protein interaction, in addition, the improvement of pharmacological properties. The potent activity profiles of the synthesized compounds suggested that dual inhibition of GSK-3β and the CXXC5-Dvl interaction could be an appropriate approach towards safely and efficientlyactivating Wntsignaling. Thus, dual-targeting inhibitors are potentially better candidates for efficient activation ofWntsignaling compared to GSK-3β inhibitors.

Keywords:  CXXC5-Dvl interaction; Dual inhibition; GSK-3β; Indirubin-3′-alkoxime derivatives; Wnt signaling; Wound healing

DOI:  https://doi.org/10.1016/j.bioorg.2022.105664
J Clin Invest. 2022 02 15. pii: e141848. [Epub ahead of print]132(4):

Age-related GSK3β overexpression drives podocyte senescence and glomerular aging.

Yudong Fang, Bohan Chen, Zhangsuo Liu, Athena Y Gong, William T Gunning, Yan Ge, Deepak Malhotra, Amira F Gohara, Lance D Dworkin, Rujun Gong.

  As life expectancy continues to increase, clinicians are challenged by age-related renal impairment that involves podocyte senescence and glomerulosclerosis. There is now compelling evidence that lithium has a potent antiaging activity that ameliorates brain aging and increases longevity in Drosophila and Caenorhabditis elegans. As the major molecular target of lithium action and a multitasking protein kinase recently implicated in a variety of renal diseases, glycogen synthase kinase 3β (GSK3β) is overexpressed and hyperactive with age in glomerular podocytes, correlating with functional and histological signs of kidney aging. Moreover, podocyte-specific ablation of GSK3β substantially attenuated podocyte senescence and glomerular aging in mice. Mechanistically, key mediators of senescence signaling, such as p16INK4A and p53, contain high numbers of GSK3β consensus motifs, physically interact with GSK3β, and act as its putative substrates. In addition, therapeutic targeting of GSK3β by microdose lithium later in life reduced senescence signaling and delayed kidney aging in mice. Furthermore, in psychiatric patients, lithium carbonate therapy inhibited GSK3β activity and mitigated senescence signaling in urinary exfoliated podocytes and was associated with preservation of kidney function. Thus, GSK3β appears to play a key role in podocyte senescence by modulating senescence signaling and may be an actionable senostatic target to delay kidney aging.

Keywords:  Aging; Bipolar disorder; Cellular senescence; Cytoskeleton; Nephrology

DOI:  https://doi.org/10.1172/JCI141848
Nat Commun. 2022 Feb 16. 13(1): 899

A non-catalytic scaffolding activity of hexokinase 2 contributes to EMT and metastasis.

Catherine S Blaha, Gopalakrishnan Ramakrishnan, Sang-Min Jeon, Veronique Nogueira, Hyunsoo Rho, Soeun Kang, Prashanth Bhaskar, Alexander R Terry, Alexandre F Aissa, Maxim V Frolov, Krushna C Patra, R Brooks Robey, Nissim Hay.

Hexokinase 2 (HK2), which catalyzes the first committed step in glucose metabolism, is induced in cancer cells. HK2's role in tumorigenesis has been attributed to its glucose kinase activity. Here, we describe a kinase independent HK2 activity, which contributes to metastasis. HK2 binds and sequesters glycogen synthase kinase 3 (GSK3) and acts as a scaffold forming a ternary complex with the regulatory subunit of protein kinase A (PRKAR1a) and GSK3β to facilitate GSK3β phosphorylation and inhibition by PKA. Thus, HK2 functions as an A-kinase anchoring protein (AKAP). Phosphorylation by GSK3β targets proteins for degradation. Consistently, HK2 increases the level and stability of GSK3 targets, MCL1, NRF2, and particularly SNAIL. In addition to GSK3 inhibition, HK2 kinase activity mediates SNAIL glycosylation, which prohibits its phosphorylation by GSK3. Finally, in mouse models of breast cancer metastasis, HK2 deficiency decreases SNAIL protein levels and inhibits SNAIL-mediated epithelial mesenchymal transition and metastasis.

DOI: https://doi.org/10.1038/s41467-022-28440-3
J Microbiol. 2022 Feb 14.

Characterization of a cold-adapted debranching enzyme and its role in glycogen metabolism and virulence of Vibrio vulnificus MO6-24/O.

Ah-Reum Han, Haeyoung Kim, Jong-Tae Park, Jung-Wan Kim.

  Vibrio vulnificus MO6-24/O has three genes annotated as debranching enzymes or pullulanase genes. Among them, the gene encoded by VVMO6_03032 (vvde1) shares a higher similarity at the amino acid sequence level to the glycogen debranching enzymes, AmyX of Bacillus subtilis (40.5%) and GlgX of Escherichia coli (55.5%), than those encoded by the other two genes. The vvde1 gene encoded a protein with a molecular mass of 75.56 kDa and purified Vvde1 efficiently hydrolyzed glycogen and pullulan to shorter chains of maltodextrin and maltotriose (G3), respectively. However, it hydrolyzed amylopectin and soluble starch far less efficiently, and β-cyclodextrin (β-CD) only rarely. The optimal pH and temperature of Vvde1 was 6.5 and 25°C, respectively. Vvde1 was a cold-adapted debranching enzyme with more than 60% residual activity at 5°C. It could maintain stability for 2 days at 25°C and 1 day at 35°C, but it destabilized drastically at 40°C. The Vvde1 activity was inhibited considerably by Cu2+, Hg2+, and Zn2+, while it was slightly enhanced by Co2+, Ca2+, Ni2+, and Fe2+. The vvde1 knock-out mutant accumulated more glycogen than the wild-type in media supplemented with 1.0% maltodextrin; however, the side chain length distribution of glycogen was similar to that of the wild-type except G3, which was much more abundant in the mutant. Therefore, Vvde1 seemed to debranch glycogen with the degree of polymerization 3 (DP3) as the specific target branch length. Virulence of the pathogen against Caenorhabditis elegans was attenuated significantly by the vvde1 mutation. These results suggest that Vvde1 might be a unique glycogen debranching enzyme that is involved in both glycogen utilization and shaping of glycogen molecules, and contributes toward virulence of the pathogen.

Keywords:  Vibrio vulnificus; cold-adapted; glycogen debranching enzyme; side chain length; virulence

DOI:  https://doi.org/10.1007/s12275-022-1507-3
Neurochem Res. 2022 Feb 15.

1-(7-Chloroquinolin-4-yl)-N-(4-Methoxybenzyl)-5-Methyl-1H-1,2, 3-Triazole-4- carboxamide Reduces Aβ Formation and Tau Phosphorylation in Cellular Models of Alzheimer's Disease.

Mariana G Fronza, Manoela Sacramento, Diego Alves, Domenico Praticò, Lucielli Savegnago.

  1-(7-Chloroquinolin-4-yl)-N-(4-methoxybenzyl)-5-methyl-1H-1,2,3-triazole-4- carboxamide (QTC-4-MeOBnE) is a new multi-target directed ligand (MTDL) rationally designed to have affinity with β-secretase (BACE), Glycogen Synthase Kinase 3β (GSK3β) and acetylcholinesterase, which are considered promising targets on the development of disease-modifying therapies against Alzheimer's Disease (AD). Previously, QTC-4-MeOBnE treatment showed beneficial effects in preclinical AD-like models by influencing in vivo neurogenesis, oxidative and inflammatory pathways. However, the biological effect and mechanism of action exerted by QTC-4-MeOBnE in AD cellular models have not been elucidated yet. Hereby we investigate the acute effect of QTC-4-MeOBnE on neuronal cells overexpressing Amyloid Protein Precursor (APP) or human tau protein, the two main features of the AD pathophysiology. When compared to the control group, QTC-4-MeOBnE treatment prevented amyloid beta (Aβ) formation through the downregulation of APP and BACE levels in APPswe-expressing cells. Furthermore, in N2a cells overexpressing human tau, QTC-4-MeOBnE reduced the levels of phosphorylated forms of tau via the modulation of the GSK3β pathway. Taken together, our findings provide new insights into the mechanism of action exerted by QTC-4-MeOBnE in AD cellular models, and further support its potential as an interesting therapeutic strategy against AD.

Keywords:  Amyloid beta; BACE; Kinases; Multi-target; Neuronal cells; Tau phosphorylation

DOI:  https://doi.org/10.1007/s11064-021-03514-8
J Ethnopharmacol. 2022 Feb 14. pii: S0378-8741(22)00145-3. [Epub ahead of print] 115107

Multi-target neuroprotective effects of herbal medicines for Alzheimer's disease.

Filipe Fernandes, Fátima Barroso, Angela De Simone, Eliška Emriková, Mónica Teixeira, José Paulo Pereira, Jakub Chlebek, Virgínia Cruz Fernandes, Francisca Rodrigues, Vincenza Andrisano, Cristina Delerue-Matos, Clara Grosso.

   ETHNOPHARMACOLOGICAL RELEVANCE: Alzheimer's disease is the most common form of dementia, but its treatment options remain few and ineffective. To find new therapeutic strategies, natural products have gained interest due to their neuroprotective potential, being able to target different pathological hallmarks associated with this disorder. Several plant species are traditionally used due to their empirical neuroprotective effects and it is worth to explore their mechanism of action.
AIM OF THE STUDY: This study intended to explore the neuroprotective potential and the safety in regard to pesticide residues of seven traditional medicinal plants, namely Scutellaria baicalensis, Ginkgo biloba, Hypericum perforatum, Curcuma longa, Lavandula angustifolia, Trigonella foenum-graecum and Rosmarinus officinalis. The safety assessment with reference to pesticides residues was also aimed.
MATERIALS AND METHODS: Decoctions prepared from these species were chemically characterized by HPLC-DAD and screened for their ability to scavenge four different free radicals (DPPH●, ABTS●+, O2●‒ and ●NO) and to inhibit enzymes related to neurodegeneration (cholinesterases and glycogen synthase kinase-3β). Cell viability through MTT assay was also evaluated in two different brain cell lines, namely non-tumorigenic D3 human brain endothelial cells (hCMEC/D3) and NSC-34 motor neurons. Furthermore, and using GC, 20 pesticides residues were screened.
RESULTS: Regarding chemical composition, chromatographic analysis revealed the presence of several flavonoids, phenolic acids, curcuminoids, phenolic diterpenoids, one alkaloid and one naphthodianthrone in the seven decoctions. All extracts were able to scavenge free radicals and were moderate glycogen synthase kinase-3β inhibitors; however, they displayed weak to moderate acetylcholinesterase and butyrylcholinesterase inhibition. G. biloba and L. angustifolia decoctions were the less cytotoxic to hCMEC/D3 and NSC-34 cell lines. No pesticides residues were detected.
CONCLUSIONS: The results extend the knowledge on the potential use of plant extracts to combat multifactorial disorders, giving new insights into therapeutic avenues for Alzheimer's disease.

Keywords:  Alzheimer's disease; Cholinesterases; Glycogen synthase kinase-3β; Oxidative stress; Pesticides residues; Plant extracts

DOI:  https://doi.org/10.1016/j.jep.2022.115107
Front Pharmacol. 2021 ;12 804615

Magnesium-Assisted Cisplatin Inhibits Bladder Cancer Cell Survival by Modulating Wnt/β-Catenin Signaling Pathway.

Tianye Li, Zihan Tang, Chunting Li, Xiaoya Liu, Linglin Cheng, Zhijing Yang, Xiaojin Zhu, Weiwei Liu, Yongye Huang.

  Magnesium, an essential mineral micronutrient, plays a role in the activation of various transporters and enzymes. The present study aimed to investigate the possibility of applying magnesium to enhance the efficacy of cisplatin which is still ranked as one of the major chemotherapeutic drugs for bladder cancer patients. Results showed that the survival rate and colony formation of bladder cancer cells were reduced by combinatorial treatment with cisplatin and magnesium chloride (MgCl2). The proportion of apoptotic cells was also increased in UC3 bladder cancer cells treated with a combination of cisplatin and MgCl2. Most importantly, a marked decrease in nuclear β-catenin was observed in cells that received cisplatin treatment. In addition, the nuclear β-catenin in cisplatin treated cells was further down-regulated by supplementing MgCl2. 6-bromoindirubin-3'-oxime (BIO), an inhibitor of glycogen synthase kinase-3 (GSK-3) that activates the Wnt/β-catenin signaling pathway by modulating β-catenin activity, was thus applied to further exploit the role of this signaling pathway in magnesium aided cancer treatment. The survival rate of bladder cancer cells was decreased by BIO treatment at concentrations of 1.0, 2.5 and 5.0 μM accompanied by increased β-catenin expression. However, the expression of β-catenin in MgCl2-treated cells was lower than in untreated cells under the same BIO concentration. The expression of cleaved caspase-3, cleaved caspase-9 and microtubule-associated protein 1 light chain 3- II (LC3-II) was highest in cells treated with MgCl2 and 5.0 μM BIO among the examined groups. Our findings reveal that magnesium could contribute to cisplatin-based chemotherapy by moderately regulating the Wnt/β-catenin signaling pathway.

Keywords:  autophagy; cancer; cisplatin; magnesium; β-catenin

DOI:  https://doi.org/10.3389/fphar.2021.804615
Front Mol Neurosci. 2021 ;14 804130

The GSK-3 Inhibitor CT99021 Enhances the Acquisition of Spatial Learning and the Accuracy of Spatial Memory.

Yeseul Lee, Zuner A Bortolotto, Clarrisa A Bradley, Thomas M Sanderson, Min Zhuo, Bong-Kiun Kaang, Graham L Collingridge.

  Glycogen synthase kinase 3 (GSK-3) is a Ser/Thr protein kinase that regulates many cellular processes, including synaptic plasticity. Previously, we reported that inhibition of GSK-3 prevents the induction of one of the major forms of synaptic plasticity, N-methyl-D-aspartate receptor (NMDAR)-dependent long-term depression (LTD), in hippocampal slices. In the present study, we have investigated the effects of inhibiting GSK-3 on learning and memory in healthy naïve animals. Systemic administration of a highly selective GSK-3 inhibitor, CT99021, reversibly blocked NMDAR-dependent LTD in the CA1 region of the hippocampus in anesthetized adult mice. In behavioral tasks, CT99021 had no effect on locomotor activity, anxiety, hippocampus-dependent contextual fear memory, and hippocampus-dependent reversal learning. However, CT99021 facilitated the rate of learning in the Morris water maze (MWM) and T-maze and enhanced the accuracy of long-term spatial memory in the MWM. These findings suggest that GSK-3 regulates the accuracy of spatial memory acquisition and recall.

Keywords:  CT99021; DMTP T-maze; GSK-3; Morris water maze; NMDAR-dependent LTD; memory enhancement; spatial learning and memory

DOI:  https://doi.org/10.3389/fnmol.2021.804130
J Therm Biol. 2022 Feb;pii: S0306-4565(21)00317-X. [Epub ahead of print]104 103149

Energy metabolism response of Litopenaeus vannamei to combined stress of acute cold exposure and waterless duration: Implications for physiological regulation and waterless live transport.

Defeng Xu, Jiaxin Wu, Lijun Sun, Xiaoming Qin, Xiuping Fan, Xiaoxian Zheng.

  Maintaining the homeostasis of energy metabolism is crucial for organism's stress tolerance and survival. Acute cold exposure (AC) and waterless duration (WD) represent the two predominate abiotic stressors during waterless live transport of Litopenaeus vannamei. Although previous reports have explored the physiological response of L. vannamei to combined stress AC + WD, the roles of energy metabolism response in regulation of stress tolerance remains unknown. The present study comparatively examined the variations of energy metabolism-related indicators in hemolymph (cortisol, hemocyanin, glucose and lactate), hepatopancreas and muscle tissues (levels of lactate and glycogen, activities of hexokinase (HK), phosphofructokinase (PFK), pyruvate kinase (PK), lactate dehydrogenase (LDH), succinate dehydrogenase (SDH) and adenosine triphosphatase (ATPase), and ATP levels). Combined stress significantly disturbed the homeostasis of energy metabolism with the increase in levels of hemocyanin, glucose and lactate, and decrease in glycogen and ATP content (P < 0.05). In addition, the activities of HK, PFK, PK, and SDH initially elevated and then decreased with the prolongation of combined stress from 3h to 9h duration, while the activity of lactate dehydrogenase (LDH) remained gradual elevation and ATPase activity decreased in a duration time dependent manner throughout the experiment. These alterations revealed that exposure to combined stress could accelerate anaerobic metabolism at initial stage and inhibit aerobic metabolism in a duration time-dependent manner, following with the reduction of energy biosynthesis and the disturbance of energy metabolism equilibrium. On the other hand, the progressive impairment on hepatopancreas tissue was observed under combined stress. In summary, the deficiency of ATP supply and histopathological injures on hepatopancreas tissue might the underlying mechanisms inducing mortality of L. vannamei during live transport.

Keywords:  Combined stress; Energy metabolism response; Hepatopancreas impairment; L. vannamei; Live transport

DOI:  https://doi.org/10.1016/j.jtherbio.2021.103149
Int J Environ Res Public Health. 2022 Jan 20. pii: 1155. [Epub ahead of print]19(3):

Effects of Perch Essence Supplementation on Improving Exercise Performance and Anti-Fatigue in Mice.

Mon-Chien Lee, Yi-Ju Hsu, Yu-Qian Lin, Ling-Ni Chen, Mu-Tsung Chen, Chi-Chang Huang.

  Silver perch (Bidyanus bidyanus) has many nutrition and health benefits, being a rich source of macro and micronutrients, phospholipids, polyunsaturated fatty acids, and a variety of essential minerals while having a high protein content. In addition to direct consumption, it is often made into a soup as an important nutritional supplement for strengthening the body and delaying fatigue. By extracting the essence, its quality can be controlled, and it is convenient to supplement. This study aimed to evaluate the effect of supplementation with Santé premium silver perch essence (SPSPE) on improving exercise performance and anti-fatigue. Fifty male institute of cancer research (ICR) mice were divided into five groups (n = 10/group): (1) vehicle (vehicle control or water only), (2) isocaloric (0.93 g casein/kg/mice/day), (3) SPSPE-1X (0.99 g/kg/mice/day), (4) SPSPE-2X (1.98 g/kg/mice/day), and (5) SPSPE-5X (4.95 g/kg/mice/day). A sample or an equal volume of liquid was fed orally for four consecutive weeks. Grip strength and swimming exhaustion tests were used as exercise performance assessments. After 10 and 90 min of unloaded swimming, biochemical parameters of fatigue were evaluated. We found that supplementation with SPSPE for four consecutive weeks could significantly improve mice's grip strength, exercise endurance performance, and glycogen content (p < 0.05), and significantly reduced post-exercise fatigue biochemical parameters, such as lactate, blood ammonia (NH3), blood urea nitrogen (BUN) concentration, and muscle damage index creatine kinase (CK) activity (p < 0.05). In summary, supplementation with SPSPE for 4 weeks could effectively improve exercise performance, reduce sports fatigue, and accelerate fatigue recovery. In addition, it did not cause any physiological or histopathological damage.

Keywords:  anti-fatigue; exercise performance; fish soup; perch essence

DOI:  https://doi.org/10.3390/ijerph19031155
Circ Res. 2022 Feb 16. CIRCRESAHA121319491

GSK-3β Localizes to the Cardiac Z-Disc to Maintain Length Dependent Activation.

Marisa J Stachowski-Doll, Maria Papadaki, Thomas G Martin, Weikang Ma, Henry M Gong, Stephanie Shao, Shi Shen, Nitha Aima Muntu, Mohit Kumar, Edith Perez, Jody L Martin, Christine S Moravec, Sakthivel Sadayappan, Stuart G Campbell, Thomas Irving, Jonathan A Kirk.

   BACKGROUND: Altered kinase localization is gaining appreciation as a mechanism of cardiovascular disease. Previous work suggests GSK-3β (glycogen synthase kinase 3β) localizes to and regulates contractile function of the myofilament. We aimed to discover GSK-3β's in vivo role in regulating myofilament function, the mechanisms involved, and the translational relevance.
METHODS: Inducible cardiomyocyte-specific GSK-3β knockout mice and left ventricular myocardium from nonfailing and failing human hearts were studied.
RESULTS: Skinned cardiomyocytes from knockout mice failed to exhibit calcium sensitization with stretch indicating a loss of length-dependent activation (LDA), the mechanism underlying the Frank-Starling Law. Titin acts as a length sensor for LDA, and knockout mice had decreased titin stiffness compared with control mice, explaining the lack of LDA. Knockout mice exhibited no changes in titin isoforms, titin phosphorylation, or other thin filament phosphorylation sites known to affect passive tension or LDA. Mass spectrometry identified several z-disc proteins as myofilament phospho-substrates of GSK-3β. Agreeing with the localization of its targets, GSK-3β that is phosphorylated at Y216 binds to the z-disc. We showed pY216 was necessary and sufficient for z-disc binding using adenoviruses for wild-type, Y216F, and Y216E GSK-3β in neonatal rat ventricular cardiomyocytes. One of GSK-3β's z-disc targets, abLIM-1 (actin-binding LIM protein 1), binds to the z-disc domains of titin that are important for maintaining passive tension. Genetic knockdown of abLIM-1 via siRNA in human engineered heart tissues resulted in enhancement of LDA, indicating abLIM-1 may act as a negative regulator that is modulated by GSK-3β. Last, GSK-3β myofilament localization was reduced in left ventricular myocardium from failing human hearts, which correlated with depressed LDA.
CONCLUSIONS: We identified a novel mechanism by which GSK-3β localizes to the myofilament to modulate LDA. Importantly, z-disc GSK-3β levels were reduced in patients with heart failure, indicating z-disc localized GSK-3β is a possible therapeutic target to restore the Frank-Starling mechanism in patients with heart failure.

Keywords:  animals; calcium; connectin; myofibrils; rats

DOI:  https://doi.org/10.1161/CIRCRESAHA.121.319491
J Stroke Cerebrovasc Dis. 2022 Feb 10. pii: S1052-3057(22)00046-5. [Epub ahead of print]31(4): 106349

Tideglusib Ameliorates Ischemia/Reperfusion Damage by Inhibiting GSK-3β and Apoptosis in Rat Model of Ischemic Stroke.

Balu Joshi, Devendra Singh, Himika Wasan, Uma Sharma, K H Reeta.

   OBJECTIVES: Glycogen synthase kinase-3β (GSK-3β), a serine/threonine protein kinase, gets activated and worsen stroke outcome after ischemia/reperfusion (I/R) injury by inducing inflammation and apoptosis. In this study, tideglusib, a selective irreversible and non-ATP competitive inhibitor of GSK-3β, was explored in cerebral I/R damage using middle cerebral artery occlusion (MCAo) model in rats.
MATERIALS AND METHODS: MCAo was done for 90 min in male Wistar rats (250-280 g) using doccol suture. In pre-treatment group, tideglusib (50 mg/kg) was administered once daily for 2 days and on the day of surgery, 30 min before MCAo. Next day, rats were examined for neurobehavioral parameters and MRI was performed to assess brain damage. In post-treatment group, tideglusib was started at 30 min after MCAo and continued for the next 2 days. After 72 h of MCAo, behavioral parameters and brain damage by MRI were assessed. Further, oxidative stress markers (MDA and GSH), inflammatory cytokines (TNF-α, IL-1β and IL-10) and expression levels of pGSK-3β S9, Bcl-2 and Bax were estimated in pre-treatment group.
RESULTS: Tideglusib pre-treatment but not post-treatment significantly improved neurobehavioral parameters (p < 0.05) and reduced brain damage (p < 0.01) when compared with MCAo group. I/R induced changes in MDA (p < 0.01), TNF-α and IL-1β (p < 0.05) were significantly attenuated by pre-treatment. Further, tideglusib pre-treatment ameliorated MCAo induced altered expressions of pGSK-3β S9, Bcl-2 and Bax.
CONCLUSION: The results of our exploratory study indicated prophylactic potential of tideglusib in I/R injury by modulating pGSK-3β S9, apoptosis and neuro-inflammation.

Keywords:  GSK-3β; Ischemia; MCAo; Reperfusion; Tideglusib

DOI:  https://doi.org/10.1016/j.jstrokecerebrovasdis.2022.106349
Cells. 2022 Jan 24. pii: 383. [Epub ahead of print]11(3):

Physiological Function during Exercise and Environmental Stress in Humans-An Integrative View of Body Systems and Homeostasis.

Gavin Travers, Pascale Kippelen, Steven J Trangmar, José González-Alonso.

  Claude Bernard's milieu intérieur (internal environment) and the associated concept of homeostasis are fundamental to the understanding of the physiological responses to exercise and environmental stress. Maintenance of cellular homeostasis is thought to happen during exercise through the precise matching of cellular energetic demand and supply, and the production and clearance of metabolic by-products. The mind-boggling number of molecular and cellular pathways and the host of tissues and organ systems involved in the processes sustaining locomotion, however, necessitate an integrative examination of the body's physiological systems. This integrative approach can be used to identify whether function and cellular homeostasis are maintained or compromised during exercise. In this review, we discuss the responses of the human brain, the lungs, the heart, and the skeletal muscles to the varying physiological demands of exercise and environmental stress. Multiple alterations in physiological function and differential homeostatic adjustments occur when people undertake strenuous exercise with and without thermal stress. These adjustments can include: hyperthermia; hyperventilation; cardiovascular strain with restrictions in brain, muscle, skin and visceral organs blood flow; greater reliance on muscle glycogen and cellular metabolism; alterations in neural activity; and, in some conditions, compromised muscle metabolism and aerobic capacity. Oxygen supply to the human brain is also blunted during intense exercise, but global cerebral metabolism and central neural drive are preserved or enhanced. In contrast to the strain seen during severe exercise and environmental stress, a steady state is maintained when humans exercise at intensities and in environmental conditions that require a small fraction of the functional capacity. The impact of exercise and environmental stress upon whole-body functions and homeostasis therefore depends on the functional needs and differs across organ systems.

Keywords:  brain; dehydration; heart; hyperthermia; lungs; muscles

DOI:  https://doi.org/10.3390/cells11030383
Front Oncol. 2021 ;11 781344

GBE1 Is an Independent Prognostic Marker and Associated With CD163+ Tumor-Associated Macrophage Infiltration in Lung Adenocarcinoma.

Yicheng Liang, Yangyang Lei, Mei Liang, Minjun Du, Zixu Liu, Xingkai Li, Xiangzhi Meng, Boxuan Zhou, Yushun Gao.

  Glycogen branching enzyme (GBE1) is a critical gene that participates in regulating glycogen metabolism. However, the correlations between GBE1 expression and the prognosis and tumor-associated macrophages in lung adenocarcinoma (LUAD) also remain unclear. Herein, we firstly analyzed the expression level of GBE1 in LUAD tissues and adjacent lung tissues via The Cancer Genome Atlas (TCGA) database. The effect of GBE1 on prognosis was estimated by utilizing TCGA database and the PrognoScan database. The relationships between the clinical characteristics and GBE1 expression were evaluated via TCGA database. We then investigated the relationships between GBE1 and infiltration of immune cells in LUAD by utilizing the CIBERSORT algorithm and Tumor Immune Estimation Resource (TIMER) database. In addition, we used a tissue microarray (TMA) containing 92 LUAD tissues and 88 adjacent lung tissues with immunohistochemistry staining to verify the association between GBE1 expression and clinical characteristics, as well as the immune cell infiltrations. We found the expression level of GBE1 was significantly higher in LUAD tissues. High expression of GBE1 was associated with poorer overall survival (OS) in LUAD. In addition, high expression of GBE1 was correlated with advanced T classification, N classification, M classification, TNM stage, and lower grade. Moreover, GBE1 was positively correlated with infiltrating levels of CD163+ tumor-associated macrophages in LUAD. In conclusion, the expression of GBE1 is associated with the prognosis and CD163+ tumor-associated macrophage infiltration in LUAD, suggesting that it has potential to be prognostic and immunological biomarkers in LUAD.

Keywords:  CD163+ tumor-associated macrophage infiltration; GBE1; LUAD; clinical characteristics; immunohistochemistry (IHC); prognosis; tissue microarray (TMA)

DOI:  https://doi.org/10.3389/fonc.2021.781344
Muscle Nerve. 2022 Feb 16.

HIGH PREVALENCE OF PARASPINAL MUSCLE INVOLVEMENT IN ADULTS WITH McARDLE DISEASE.

R Álvarez-Velasco, C A Nuñez-Peralta, J Alonso-Pérez, E Gallardo, R Collet-Vidiella, D Reyes-Leiva, E Pascual-Goñi, L Martín-Aguilar, M Caballero-Ávila, A Carbayo-Viejo, J Llauger-Roselló, J Díaz-Manera, M Olivé.

   INTRODUCTION/AIMS: Very few studies analyzing the pattern of muscle involvement in magnetic resonance imaging (MRI) of patients with McArdle disease have been reported to date. We aimed to examine the pattern of muscle fat replacement in patients with McArdle disease.
METHODS: We performed a retrospective study including all patients with genetically confirmed McArdle disease followed in our center from January 2010 to March 2021. Clinical data were collected from the medical record. Whole-body MRI was performed as part of the diagnostic evaluation. The distribution of muscle fat replacement and its severity were analyzed.
RESULTS: Nine patients were included. Median age at onset was 7 years (range:5-58) and median age at the time when MRI was performed was 57.3 years (range 37.2-72.8). At physical examination 4 patients had permanent weakness: in 3 the weakness was limited to paraspinal muscles whereas in one the weakness involved the paraspinal and proximal upper limb muscles. Muscle MRI showed abnormalities in 6 of the 7 studied patients. In all of them fat replacement of paravertebral muscles was found. Other muscles frequently affected were the tongue in 3, subscapularis in 3, and long head of biceps femoris and semimembranosus in 2.
DISCUSSION: Our findings suggest that paraspinal muscle involvement is common in McArdle disease and support the need to include this disease in the differential diagnosis of the causes of paraspinal muscle weakness. Involvement of the tongue and subscapularis are also frequent in McArdle disease. This article is protected by copyright. All rights reserved.

Keywords:  McArdle disease; axial myopathy; glycogen storage disease; glycogen storage disease type V; magnetic resonance imaging; paraspinal muscles

DOI:  https://doi.org/10.1002/mus.27523
Nanoscale. 2022 Feb 18.

Triggering the nanophase separation of albumin through multivalent binding to glycogen for drug delivery in 2D and 3D multicellular constructs.

Agata Radziwon, Sukhvir K Bhangu, Soraia Fernandes, Christina Cortez-Jugo, Robert De Rose, Brendan Dyett, Marcin Wojnilowicz, Petra Laznickova, Jan Fric, Giancarlo Forte, Frank Caruso, Francesca Cavalieri.

Engineered nanoparticles for the encapsulation of bioactive agents hold promise to improve disease diagnosis, prevention and therapy. To advance this field and enable clinical translation, the rational design of nanoparticles with controlled functionalities and a robust understanding of nanoparticle-cell interactions in the complex biological milieu are of paramount importance. Herein, a simple platform obtained through the nanocomplexation of glycogen nanoparticles and albumin is introduced for the delivery of chemotherapeutics in complex multicellular 2D and 3D systems. We found that the dendrimer-like structure of aminated glycogen nanoparticles is key to controlling the multivalent coordination and phase separation of albumin molecules to form stable glycogen-albumin nanocomplexes. The pH-responsive glycogen scaffold conferred the nanocomplexes the ability to undergo partial endosomal escape in tumour, stromal and immune cells while albumin enabled nanocomplexes to cross endothelial cells and carry therapeutic agents. Limited interactions of nanocomplexes with T cells, B cells and natural killer cells derived from human blood were observed. The nanocomplexes can accommodate chemotherapeutic drugs and release them in multicellular 2D and 3D constructs. The drugs loaded on the nanocomplexes retained their cytotoxic activity, which is comparable with the activity of the free drugs. Cancer cells were found to be more sensitive to the drugs in the presence of stromal and immune cells. Penetration and cytotoxicity of the drug-loaded nanocomplexes in tumour mimicking tissues were validated using a 3D multicellular-collagen construct in a perfusion bioreactor. The results highlight a simple and potentially scalable strategy for engineering nanocomplexes made entirely of biological macromolecules with potential use for drug delivery.

DOI: https://doi.org/10.1039/d1nr08429a